Fig. 3: HSV-1-induced apoptosis of MEFs does not require the caspase-8 adaptor/activator FADD. | Cell Death & Differentiation

Fig. 3: HSV-1-induced apoptosis of MEFs does not require the caspase-8 adaptor/activator FADD.

From: Caspase-8 is required for HSV-1-induced apoptosis and promotes effective viral particle release via autophagy inhibition

Fig. 3

A, B Anti-caspase-8 and C anti-caspase-3 western blot analysis of total extracts of MEFs either expressing a scrambled shRNA (shCtrl) or a shRNA against FADD (shFADD), either mock-infected (0 h) or infected with HSV-1 for 8, 14 and 24 h (A, C) or treated with 50 ng/ml FasL for 4 h (B). The extent and kinetic of caspase-8 processing to the active p43/p41 and p18/p10 fragments and the processing of caspase-3 to the active p17 fragment are similar in shCtrl and shFADD MEFs. β-Actin served as a loading control. D Percentage of FITC-Annexin-V/7-AAD-negative (surviving) shCtrl and shFADD MEFs, either mock-infected (0 h) or infected with HSV-1 for 14, 24 and 48 h showing that the kinetics of HSV-1-induced apoptosis is not largely affected by the absence of FADD. E Anti-FADD western blot analysis showing the efficient knockdown of FADD in MEFs and U937 cells stably expressing FADD shRNAs (shF). As a control, a scrambled shRNA was transduced (shC). β-actin served as a loading control. F Caspase-8 (IETDase) activity assay of total extracts of shCtrl and shFADD U937 cells infected with 50 m.o.i. of HSV-1 for 0, 8, 14 and 24 h showing a similar extent and kinetic of caspase-8 activation between the two cell lines. Data in D and F represent the means of 3–6 independent experiments ± SD. Statistical evaluation by one-way ANOVA: **p < 0.01; ***p < 0.001.

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